Objective To investigate the correlation between the variations of mitochondrial gene ATP6 and type 2 diabetes mellitus ( T2DM ) and chronic complications. Methods Genomic DNA were extracted from 254 T2DM patients and 165 age-matched controls. After amplification of ATP6 by PCR and direct sequencing, all sequences were compared with the reference sequence (rCRS) to find out the variations. Bioinformatics and statistic method were used to analyze these variations. Results Many variations were detected respectively in T2DM patients and controls, a part of them only appeared in T2DM patients in low frequency, which has not been reported previously. Most of these variations are located in thethird and forth transmembrane helix of ATP synthase subunit 6 (ATPase6). Interestingly, these variationsalmost were detected in the non-obese T2DM patients with hypotension, including G8557A, A8563G,T8594C, C8609T, A8689G, G8998A and G9139A. Conclusions There were many variations in geneATP6 and must of them are mitochondrial SNP, while variations A8689G, T8825C, G8920A, G8998A andG9139A may be mild mutations which my increase the susceptibility of T2DM. G8557A, A8563G,T8594C, C8609T, A8689G, G8998A and G9139A may be associated with the biogenetics diseases suchdiabetes and hypertension.

Objective: To construct an adenovirus-mediated anti-sense RNA targeting K-ras exon 1 of SW1990 cell line and observe its effect on cell proliferation and apoptosis after transferred into SW1990 cell line. Methods: K-ras exon 1 cDNA was cloned into shuttle vector pShuttle-CMV and the resultant plasmid was confirmed by enzyme digestion and PCR. Clones with inverted insertion were selected and co-transferred into E. coli BJ5183 with an adenoviral backbone plasmid pAdEasy-1 to produce recombinant plasmid by homologous recombination. Recombinants were then selected and transfected into 293 cell line to produce recombinant adenovirus. Recombinant adenovirus production was confirmed by PCR analysis and was amplified and purified; the virus titer was determined. Ad-LacZ was used to infect SW1990 cells and the infection efficiency was observed by X-gal staining. SW1990 cells was infected with the recombinant adenovirus and their proliferation and apoptosis were determined by MTT and annexin V/PI FCM assay. Results: A 282 bp target gene fragment was acquired by PCR; the titer of recombinant adenovirus was 7.6 × 108 pfu/ml before purification by CsCl2 gradient centrifugation and 5.0 × 1910 pfu/ml after CsCl2 gradient centrifugation. When the recombinant adenovirus was at 100 MOI, the infection efficiency of SW1990 cells nearly reached 100%. The transfection of recombinant adenovirus significantly inhibited SW1990 cell proliferation (P<0.05), with a maximal inhibitory rate of 40.5% 4-5 days after infection; it also significantly increased SW1990 cell apoptosis, with the apoptotic rate being (22.54 ± 5.38) % 72 hours after infection. Conclusion: We have successfully constructed an anti-sense RNA adenovirus vector targeting K-ras exon 1 of SW1990 cell line, paving a way for the anti-sense K-ras gene therapy of pancreatic carcinoma.

For the purposes of meeting the needs of COPD patients for health education, decreasing times and costs of re-hospitalization, and increasing patient satisfaction and nurses' working enthusiasm. In 2011, our hospital established a specialist nursing clinic for chronic obstructive pulmonary disease (COPD), and two nursesentitled-deputy directorswith rich clinical experiences for more than fifteen years and strong communication skills were se-lected to take charge of the clinic, and they were responsible for nursing assessment, making individualized pul-monary rehabilitation plan, health guidance, long-term follow up, etc., for discharged stable COPD patients and COPD patients with no need for hospitalization. Patient satisfaction was improved from 83% to 96% after implemen-tation.

Child ; Developmental Disabilities ; Ethics, Medical ; Fertilization in Vitro ; adverse effects ; Humans ; Treatment Outcome

Objective To evaluate the effect of remifentanil on cell apoptosis during renal ischemia-reperfusion (I/R) in rats.Methods Seventy-five male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =25 each):sham operation group (group S),I/R group,and remifentanil group (group R).Renal ischemia was induced by occlusion of the bilateral renal arteries for 45 min followed by reperfusion in groups I/R and R.Remifentanil was infused at 1.0 μg· kg-1 · min-1 via the caudal vein starting from 15 min before ischemia until 30 min of reperfusion in group R,while the equal volume of normal saline was given instead of remifentanil in groups S and I/R.At 15 min before ischemia (T0) and 3,6,12,24 h of reperfusion (T1-4),5rats were anesthetized and sacrificed,and renal specimens were obtained to detect the apoptotic rate and expression of Bax and Bcl-2 protein (by flow cytometry) and mRNA (by RT-PCR).The ratios between Bcl-2/Bax protein and mRNA expression were calculated.The pathological changes of renal tubules were scored.Results Compared with group S,the pathological scores and apoptotic rate were significantly increased at T1-4,and ratios between Bcl-2/Bax protein and mRNA expression were increased at T1,2,while decreased at T3,4 in groups R and I/R (P ＜0.01).Compared with group I/R,the pathological scores and apoptotic rate were significantly decreased at T1-4,while the ratios between Bcl-2/Bax protein and mRNA expression were increased in group R (P ＜ 0.05 or 0.01).Compared with the baseline value at T0,the pathological scores and apoptotic rates were significantly increased at T1 4,and the ratios of Bcl-2/Bax protein and mRNA expression were increased at T1,2,while decreased at T3,4 in groups R and I/R (P ＜ 0.01).Conclusion Regulation of Bcl-2/Bax expression and inhibition of cell apoptosis in renal tissues are involved in the mechanism by which remifentanil reduces renal I/R injury in rats.

Objective To explore the effects of lipoxinA4 on expression of cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2) in rat primary lung fibroblast cells (LF) after lipopolysaccharide (LPS) challenge.Methods Primary lung fibroblast cells were incubated with various concentrations (0.1,1,10 μg/mL) of LPS for different lengths of time (3,6,9 h).Then primary lung fibroblast cells were still incubated in DMEM medium containing LPS in the presence or absence of lipoxinA4.After incubation,the supematant of medium was collected and the level of PGE2 was detected by using ELISA.The cells were harvested,and COX-2 protein was analyzed by Western blot.Results The model of acute inflammation in fibroblasts was well established by administering 1 μg/mL LPS in fibroblasts for 6 hours.Induction of COX-2 protein by LPS was inhibited by lipoxinA4.The levels of PGE2 in control group,LPS group and LPS + LipoxinA4 group were 55.84 pg/mL,411.73 pg/mL and 307.07 pg/mL,respectively,and there was a significantdifference between LPS group and LPS + LipoxinA4 group (P ＜0.01).Conclusion LipoxinA4 down-regulates the expression of the COX-2 induced by LPS in primary lung fibroblast cells and consequently inhibits the production of PGE2 in a dose dependent manner.

Objective To investigate the role of opioid receptors in remifentanil-induced attenuation of renal ischemia/reperfusion (I/R) injury in rats.Methods Seventy-five male Sprague-Dawley rats weighing 250-300 g were randomly divided into 5 groups ( n =15 each):sham operation group (group S),group I/R,remifentanil group (group R),naloxone group (group N),and naloxone + remifentanil group (group NR).Renal ischemia was induced by clamping the bilateral renal arteries for 45 min using an atraumatic clamp followed by reperfusion.In groups R and NR,remifentanil was infused at 1.0 μg· kg-1 · min-1 via the caudal vein starting from 15 min before ischemia until 30 min of reperfusion,while groups S,I/R and N received the equal volume of normal saline instead of remifentanil.In groups N and NR,naloxone 0.3 mg/kg was injected via the caudal vein at 20 min before ischemia and at 35 min after ischemia respectively,while groups S,I/R and R received the equal volume of normal saline instead of naloxone.Blood and urine samples were collected from the femoral vein and urinary bladder respectively at 24 h of reperfusion for determination of the levels of serum creatinine (Cr) and blood urea nitrogen (BUN),urinary N-acetyl-β-D-glucosaminidase (NAG) and γ-glutamyl transpeptidase (γ-GT).The rats were sacrificed at 24 h of reperfusion and the renal tissues were removed for determination of nalondialdehyde (MDA) content and superoxide dismutase (SOD) activity.Pathological changes in renal tissues were observed with light microscope.Results Compared withgroup S,the levels of serum Cr and BUN,urinary NAG and γ-GT,and MDA were significantly increased,while the activity of SOD was significantly decreased in the other 4 groups ( P ＜ 0.05 or 0.01 ) and pathological changes in renal tissues were observed in the other 4 groups.Compared with group I/R,the levels of serum Cr and BUN,urinary NAG and γ-GT levels,and MDA were significantly decreased,while the activity of SOD was significantly increased ( P ＜ 0.01 ),and the pathological changes were reduced in group R,and no significant change was found in the parameters mentioned above in groups N and NR ( P ＞ 0.05).The pathological changes were similar in groups I/R,N and NR.Compured with group R,serum Cr and BUN concentrations,urinary NAG and γ-GT levels and MDA concent were increased,while SOD activity were decreased ( P ＜ 0.05 or 0.01 ).Conclusion Opioid receptors mediate remifentanil-induced attenuation of renal I/R injury in rats.

Objective To investigate the protection effects of teprenone in aspirin-induced gastric mucosa injury.Methods From 2008 to 2010,a total of 296 patients who took aspirin for the first time at the Department of Cardiovascular,First Hospital Affiliated to Zhejiang Chinese Medicine University were randomly divided into two groups.There were 166 cases in aspirin group,which took aspirin 100mg daily; 130 cases in aspirin and teprenone group,the aspirin dose equivalent with aspirin group and took teprenone 50mg/time,3 times/day orally.Gastrointestinal symptoms and gastric mucosa injury of patients in these two group were inspected at 3 month,6 month and 1 year.Results A total of 143 cases were recruited in aspirin group and 118 cases in aspirin and teprenone group.After taking medicine for 3 months,the occurrence rate of gastrointestinal symptoms in aspirin group was 1.40 ％.Compared with aspirin and teprenone group,the difference was statistical significant (0,x2 =1.663,P＝ 0.197).Follow up after taking medicine for 6 months,the occurrence rate of gastrointestinal symptoms in aspirin group was 4.96％.Compared with aspirin and teprenone group,the difference was statistical significant (0,x2 ＝6.021,P＝0.014).Follow up after taking medicine for 1 year,the occurrence rate of gastrointestinal symptoms in aspirin group was 20.15 ％.Compared with aspirin and teprenone group,the difference was statistical significant (1.69％,x2 =20.984,P=0.001).Compared with aspirin group,the symptom and endoscopy score of aspirin and teprenone group decreased significantly at follow-up for 6 months and 1 year (P＜0.05; P＜0.01 ).Compared with at 6 month,the symptom and endoscopy score of aspirin group at 1 year increased significantly (P＜0.05 ; P＜0.01).Conclusion Teprenone has certain protection effects in aspirin-induced gastric mucosa injury.Long-term use of conventional doses of aspirin may cause vary degrees of gastric mucosal injury,and the gastric mucosal injury get more severe as the time of taking medicine increases.

Objective To explore the influence of structured lipid emulsion on inflammatory cell factors and immunologic function in gastrointestinal cancer patients after operation.Methods From July 2010 to February 2012,80 postoperative patients with gastrointestinal cancer were randomly divided into 2 groups with 40 cases each by stratified sampling method:structured lipid emulsion group (observation group),physical mixture of medium-chain/long-chain triglycerides group (control group).Both groups received isonitrogenous and isocaloric parenteral nutrition for 6 days after operation.The general conditions after operation were compared between two groups.Inflammatory cell factors and immunologic function were measured before operation,the 1st day and 7th day after operation.Results After 6 days parenteral nutrition,the vital sign data,blood routine,liver and renal function,blood fat etc was normal in two groups.There was no significant difference in the incidence of postoperative infection complication and hospitalization duration between observation group and control group [12.5％(5/40) vs.25.0％(10/40),[( 12.9 ± 0.7 d) vs.(13.1 ±0.9) d] (P ＞0.05).The serum levels of interleukin (IL)-1,IL-2,IL-6,tumor necrosis factor (TNF)-α on the 1st day after operation in two groups decreased,then gradually increasing.The serum levels of IL-2,IL-6,TNF- α on the 7th day after operation in observation group[(987 ± 203),(277 ± 107),( 197 ± 98) ng/L] were lower than those in control group [(1347 ± 195),(752 ± 187),(342 ± 106) ng/L](P＜0.05 ).The serum levels of IgG,IgA,IgM,CD3+,CD4+ and CD8+ on the 1st day after operation in two groups decreased,then gradually increasing.The serum levels of IgG,CD3+,CD4+ were higher in observation group [( 14.2 ± 1.8 ) g/L,0.746 ± 0.111,0.580 ± 0.067] than those in control group [( 12.1 ± 2.8) g/L,0.606 ±0.101,0.404 ± 0.072] (P ＜0.05).Conclusion Structured lipid emulsion can reduce immunological suppression and inflammation in gastrointestinal cancer patients after operation,regulate the inflammatory cell factors producing and releasing,improve the immunologic function.

AIM To study the protective effects of Jingqi Capsules (Cordycps Militaris,Rehmanniae Radix Praeparata,Lycii Fructus,) on the damage to rat reproductive system induced by cyclophosphamide.METHODS After low and high dose groups of Jingqi Capsules and positive group were administered for two weeks in advance,cyclophosphamide [25 mg/(kg · d)] modeling was conducted consecutively by intraperitoneal injection for five days except the normal group,while the treatment groups were administered consecutively.All the rats were executed the day after the end of the modeling.Serum testosterone content was measured by radioimmunoassay;body weight,testis,epididymis and prostate of rats were weighed and organ coefficient was calculated;morphology of testis was observed by HE;sperm count and sperm motility were detected by CASA;apoptosis of cells was observed by TUNEL.RESULTS Compared with the model group,each dose group of Jingqi Capsules increased in the level of serum testosterone and organ coefficient of testis,epididymis and prostate.The damage to seminiferous epithelium relieved and the number of all spermatogenic cells increased.Sperm count and sperm motility increased.Decrease in apoptosis of cells was demonstrated by TUNEL.CONCLUSION Jingqi Capsules has protective effects on damage to rat reproductive system induced by cyclophosphamide.